Product info: Apa I


Name
Apa IBlocked by   
 GGGCC(5mC) methylation  
Cat. #E019E020E019TE020TE019m
Package, u.a.1000500010005000500
Concentration, u.a./ml10000-3000010000-3000010000-3000010000-3000010000

Recognition site
GGGCCC
CCCGGG
SourceAcetobacter pasteurianus
Assayed onLambda DNA (dam-dcm-, BamHI-digest)
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-dcm-, BamHI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Optimal SE-bufferY (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Enzyme activity (%)
BGOWYRose
50 - 7525 - 500 - 100 - 1010050
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationsAfter 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme 10 X SE-buffer Y, BSA (except E019m, E019T and E020T).
For E019T and E020T an additional 10X SE-buffer ROSE is supplied.
Methylation sensitivityBlocked by overlapping Dcm methylation(CmCWGG): GGGCCCWGG
Inactivation 20 minutes under
65oC
NotesTo obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Seurinck, J., Van de Voorde, A., Van Montagu, M. Nucleic Acids. Res.11: 4409-4415 (1983).
Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:

To view the fragments length values please point mouse cursor over diagram
Fragment lengths
M12345M

M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322